Journal: bioRxiv

Article Title: Antibody Transcytosis and Neutralizing Activity in Respiratory Epithelial Cells

doi: 10.64898/2026.05.25.727697

Figure Lengend Snippet: A) Immunoblot of FcRn and pIgR in hNECs and hBECs. B, C) Relative protein expression levels by Western blotting of FcRN, pIgR, normalized to β-tublin, n = <3. D, E) Quantification of flow cytometry cell types by percentage of total cells, values are expressed as mean ± SE, n = 1/group. F, G) Transcytosis of IgG and IgA was determined using an ELISA assay. Values are expressed as mean ± SE, n = <3well/time point. *P<0.05 **P<0.01

Article Snippet: Human nasal epithelial cells (hNECs) or bronchial epithelial cells (hBECs) (Promocell) were grown to confluence in 24-well Falcon filter inserts (0.4-uM pore; 0.33cm 2 ; Becton Dickinson) using PneumaCultTM-Ex Plus Medium (Stemcell, Cat# 05001).

Techniques: Western Blot, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Microfluidic analysis reveals ROCK2 regulation of endothelial cilia is essential for blood vessel lumen formation and vascular integrity

doi: 10.64898/2026.05.27.728336

Figure Lengend Snippet: (A) Schematic of a primary cilium, depicting its core compartments (basal body, transition zone, axoneme, ciliary tip) and the localization of key ciliary proteins ARL13B and RPGRIP1L. Intraflagellar transport (IFT) complexes (IFT-A and IFT-B) facilitate cargo trafficking along the axoneme. Adapted from Smith et al. (2020). (B, C) Immunofluorescence images confirming primary cilia (ARL13B, red) in human cerebral microvascular endothelial cells (hCMEC/D3) (B) and mouse bEnd.3 cells (C) . Nuclei are counterstained with DAPI (blue). Scale bars, 50 µm. (D) Primary cilia (ARL13B, red) in a monolayer of human umbilical vein endothelial cells (HUVECs). Nuclei are stained with DAPI (blue). Scale bar, 50 µm. (E) Primary cilia (ARL13B, red) on endothelial tubules (white arrowheads) and fibroblasts (yellow arrowheads) in a HUVEC–human dermal fibroblast (HDF) organotypic coculture. L, lumen. Nuclei are stained with DAPI (blue). Scale bar: 15 µm. (F, G) Endothelial primary cilia in E13.5 mouse embryonic brain vessels. Cilia (arrows, ARL13B, red) are visible on endothelial cells lining the lumen of a superficial vessel (F) and on intraparenchymal vessels (G) . Endomucin (green (F), brown (G)) marks endothelial cells; nuclei are stained with DAPI (blue). Scale bars, 50 µm (F) and 10 µm (G) .

Article Snippet: The following cell lines were cultured in a humidified atmosphere at 37°C with 5% CO 2 . hCMEC/D3 (human cerebral microvascular endothelial cells) (ATCC® CRL-3245TM) were cultured in EndoGRO-MV Complete Media (Merck, SCME004) supplemented with 1 ng/ml basic Fibroblast Growth Factor (bFGF; PeproTech, 100-18B) and used between passages 30-35 for ciliogenesis assays. bEnd.3 (mouse brain endothelial cells) (ATCC® CRL-2299TM) were cultured in DMEM (Gibco, 11995065) supplemented with 10% (v/v) heat-inactivated foetal bovine serum (FBS) (Merck, F9665), 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM L-glutamine (all from Gibco). hTERT RPE-1 (human retinal pigment epithelial cells) (ATCC® CRL-4000TM) were cultured in DMEM/F12 (Gibco, 11330032) supplemented with 10% FBS, penicillin/streptomycin, and L-glutamine.

Techniques: Immunofluorescence, Staining

Journal: PLOS One

Article Title: Scaffold-free 3D-cell co-culture model system for the study of metastatic cancer in the brain TME

doi: 10.1371/journal.pone.0349061

Figure Lengend Snippet: Top panel : HBEC-5i/SK-OV-3 (turbo GFP/green) model; Bottom panel : HBEC-5i/MDA-MB-231 (turbo FP602/red) model. Cells were seeded in equal proportion (200,000 each, ~ 30% confluence) in a 25 cm 2 culture flask and allowed to evolve for ~30 days. Self-organization of endothelial cells in 3D aggregates and accumulation of cancer cells around these structures was observable after ~7 days (80-90% confluence). After ~21 days, clearly delineated 3D networked constructs incorporating the vast majority of cancer cells were observable.

Article Snippet: HBEC-5i human brain endothelial cells were obtained from ATCC (Manassas, VA).

Techniques: Construct

Journal: PLOS One

Article Title: Scaffold-free 3D-cell co-culture model system for the study of metastatic cancer in the brain TME

doi: 10.1371/journal.pone.0349061

Figure Lengend Snippet: Top panel : HBEC-5i/SK-OV-3 (turbo GFP/green) model; Bottom panel : HBEC-5i/MDA-MB-231 (turbo FP602/red) model. Cells were seeded in a proportion of HBEC-5i (180,000 cells)/cancer cells (~20,000) in a 25 cm 2 culture flask and allowed to evolve for ~30 days. Self-organization of endothelial cells in 3D aggregates and accumulation of cancer cells in these structures was observable after ~7 days. After ~21 days, clearly delineated 3D networked constructs were observable.

Article Snippet: HBEC-5i human brain endothelial cells were obtained from ATCC (Manassas, VA).

Techniques: Construct

Journal: PLOS One

Article Title: Scaffold-free 3D-cell co-culture model system for the study of metastatic cancer in the brain TME

doi: 10.1371/journal.pone.0349061

Figure Lengend Snippet: Top panel : HBEC-5i/SK-OV-3 (turbo GFP/green) model; Bottom panel : HBEC-5i/MDA-MB-231 (turbo FP602/red) model. HBEC-5i cells (~200,000) were seeded in a 25 cm 2 culture flask and allowed to evolve until they formed 3D networked structures (~10 days), time when ~200,000 cancer cells were added to the flask. The migration of SK-OV-3 and MDA-MB-231 cells toward HBEC-5i structures was observable already after 2 days of co-culture. The evolution of endothelial/cancer constructs was monitored for an additional 3 weeks.

Article Snippet: HBEC-5i human brain endothelial cells were obtained from ATCC (Manassas, VA).

Techniques: Migration, Co-Culture Assay, Construct

Journal: PLOS One

Article Title: Scaffold-free 3D-cell co-culture model system for the study of metastatic cancer in the brain TME

doi: 10.1371/journal.pone.0349061

Figure Lengend Snippet: (A,D) Control wells with red fluorescent SK-BR-3 cells seeded in the upper inserts; ( B,C,E,F ) Experimental wells with red fluorescent SK-BR-3 cells seeded in the upper inserts and HBEC-5i in the bottom wells; (G,J) Control wells green fluorescent SK-OV-3 cells seeded in the upper inserts; ( H,I,K,L ) Experimental wells with green fluorescent SK-OV-3 cells seeded in the upper inserts and HBEC-5i in the bottom wells. The images were taken from the bottom wells after 5 days incubation.

Article Snippet: HBEC-5i human brain endothelial cells were obtained from ATCC (Manassas, VA).

Techniques: Control, Incubation

Journal: PLOS One

Article Title: Scaffold-free 3D-cell co-culture model system for the study of metastatic cancer in the brain TME

doi: 10.1371/journal.pone.0349061

Figure Lengend Snippet: Top panel: HBEC-5i/SK-OV-3 (turbo GFP/green) model; (A) Green fluorescent SK-OV-3 cells (FITC filter); (B) Dead cells (red) visualized by PI staining (TRITC filter); (C) Merged image of transmitted, FITC- and TRITC-filtered images of co-cultured cells. Bottom panel: HBEC-5i/MDA-MB-231 (turbo FP602/red) model; (D) Red fluorescent MDA-MB-231 cells (TRITC filter); (E) Dead cells (green) visualized by staining with EasyProbe dye (FITC filter); (F) Merged image of transmitted, TRITC- and FITC-filtered images of co-cultured cells. Images were acquired after 28 days of co-culture.

Article Snippet: HBEC-5i human brain endothelial cells were obtained from ATCC (Manassas, VA).

Techniques: Staining, Cell Culture, Co-Culture Assay

Journal: PLOS One

Article Title: Scaffold-free 3D-cell co-culture model system for the study of metastatic cancer in the brain TME

doi: 10.1371/journal.pone.0349061

Figure Lengend Snippet: Top panel : monoculture controls of HBEC-5i and SK-BR-3 cells treated with Lapatinib (10 µM); (A,B) Dead cells (green) in HBEC-5i monocultures; (C,D) Dead cells (yellow) in red fluorescent SK-BR-3 monocultures. Bottom panel : Co-culture HBEC-5i/SK-BR-3 (turbo FP602/red) cell model treated with Lapatinib (1 and 10 µM). (E-H) Dead cells (yellow) in the endothelial/cancer co-culture system. EasyProbe green dye (FITC filter) was used to visualize the dead cells in all cultures.

Article Snippet: HBEC-5i human brain endothelial cells were obtained from ATCC (Manassas, VA).

Techniques: Co-Culture Assay

Journal: Pharmaceuticals

Article Title: Theranostic vNAR-Based Immunoconjugates Achieve Selective Intracellular Cisplatin Delivery in Embedded 3D HER2-Positive Breast Cancer In Vitro Model

doi: 10.3390/ph19040633

Figure Lengend Snippet: Confocal microscopy analysis of vNAR FITC binding and intracellular uptake was conducted via immunofluorescence using three cell lines with defined molecular profiles: ( a ) U87-MG cells (EGFRvIII + /wtEGFR + /HER2 + ) served as the positive control for EGFRvIII recognition; ( b ) HBEC-5i cells (EGFRvIII − /wtEGFR + /HER2 − ) served as the negative control for EGFRvIII and HER2, and as the positive control for wtEGFR expression; and ( c ) SKBR3 cells (EGFRvIII + /wtEGFR + /HER2 + ) were used to assess vNAR FITC specificity for EGFRvIII in the context of co-expression of all three markers. Cells were incubated with vNAR FITC (0.081 µM) for 24 h. Immunofluorescence corresponding to vNAR FITC is indicated by green arrows, and nuclei were counterstained with propidium iodide (red). Images were acquired at 40× magnification. Scale bars: 17.35 µm ( a ), 50.5 µm ( b ), 22.14 µm ( c ). All experiments were performed in triplicate.

Article Snippet: U87-MG (ATCC HTB-14), HBEC-5i (ATCC CRL-3245), and SKBR3 (ATCC HTB-30) cells were cultured in their respective media according to ATCC guidelines and maintained at 37 °C in a 5% CO 2 atmosphere.

Techniques: Confocal Microscopy, Binding Assay, Immunofluorescence, Positive Control, Negative Control, Expressing, Incubation

Journal: Pharmaceuticals

Article Title: Theranostic vNAR-Based Immunoconjugates Achieve Selective Intracellular Cisplatin Delivery in Embedded 3D HER2-Positive Breast Cancer In Vitro Model

doi: 10.3390/ph19040633

Figure Lengend Snippet: Immunofluorescence analysis of anti-HER2 peptide of FITC-labeled C9P binding and internalization. ( a ) U87-MG cells (EGFRvIII + /wtEGFR + /HER2 + ), ( b ) HBEC-5i cells (EGFRvIII − /wtEGFR + /HER2 − ) as negative control, and ( c ) SKBR3 cells (EGFRvIII + /wtEGFR + /HER2 + ) as positive control for HER2 expression. Cells were incubated with C9P FITC (5.62 µM) for 24 h. Immunofluorescence of C9P is depicted in green and indicated by the green arrows, and nuclei were counterstained with propidium iodide (red). Magnification 40×. Scale bars: 21.08 µm ( a ), 50.5 µm ( b ), 24.3 µm ( c ). Experiments were performed in triplicate.

Article Snippet: U87-MG (ATCC HTB-14), HBEC-5i (ATCC CRL-3245), and SKBR3 (ATCC HTB-30) cells were cultured in their respective media according to ATCC guidelines and maintained at 37 °C in a 5% CO 2 atmosphere.

Techniques: Immunofluorescence, Labeling, Binding Assay, Negative Control, Positive Control, Expressing, Incubation

Journal: Glycobiology

Article Title: Hypofucosylation promotes pertussis toxin binding to cell surface glycococonjugates and pertussis toxin-induced intracellular ERK signaling

doi: 10.1093/glycob/cwag011

Figure Lengend Snippet: Inhibition of fucosylation in HBEC-3KT cells results in increased PT binding. A) Schematic of N-linked glycan synthesis inhibition in HBEC-3KT cells using small molecule inhibitors of glycosylation. 3FaxNeu5Ac and 2F-Fuc also impact other classes of glycoconjugates. HBEC-3KT cells were cultured in the presence of the inhibitors for 72 h. B) Flow cytometry analysis of PT binding to cell surfaces of the HBEC-3KT cells cultured with DMSO, 300 nM kifunensine, 50 μM 3F ax Neu5Ac, or 200 μM 2F-Fuc. Bar graph shows the quantification of geometric mean fluorescence from 3 independent trials, normalized to the geometric mean fluorescence of DMSO-treated cells. Statistical analyses were performed by one-way ANOVA (error bar indicates mean ± SD, * * * * indicates p value <0.0001). C) PT lectin blot of lysates of DMSO-, kifunensine-, 3F ax Neu5Ac-, or 2F-Fuc-treated HBEC-3KT cells. The blot shown is representative of 3 biological replicates.

Article Snippet: HBEC-3KT cells were obtained from John Minna (UT Southwestern Medical Center). ( ) Colo205 and Jurkat cells were obtained from the ATCC.

Techniques: Inhibition, Binding Assay, Glycoproteomics, Cell Culture, Flow Cytometry, Fluorescence

Journal: Nature Communications

Article Title: AI-guided multi-omics analysis identifies NPC1-modulated susceptibility to SARS-CoV-2 infection under PM 2.5 exposure

doi: 10.1038/s41467-026-71196-3

Figure Lengend Snippet: Verification of angiotensin-converting enzyme 2 (ACE2) inhibition using a series of DX600 concentrations and the corresponding viral infection efficiency in wild-type A549 (alveolar basal epithelial adenocarcinoma cells) ( a ), rs1788783 knockout (KO) A549 ( b ), primary human bronchial epithelial cells (HBECs) ( c ), and BEAS-2B (human bronchial epithelial cells) ( d ), respectively. Relative luminescence intensities are presented as mean ± standard deviation ( n = 10) from 10 biological replicates. Exact P values are shown in the figure. Reduced NPC1 expression levels using 3 distinct small interfering RNAs (siRNAs) and negative control siRNA (NC siRNA), and the corresponding viral infection efficiency in primary human bronchial epithelial cells (HBECs) ( e ) and BEAS-2B (human bronchial epithelial cells) ( f ), respectively. Viral infection was quantified by the expression levels of open reading frame 1 (ORF1) and nucleocapsid (N) genes. Data are presented as mean ± standard deviation ( n = 3) from 3 biological replicates. Statistical significance was assessed using two-sided t-tests. Source data are available in the file.

Article Snippet: Primary human bronchial epithelial cells (HBECs) were provided by Procell Life Science & Technology Co., Ltd (China, #CP-H009).

Techniques: Inhibition, Infection, Knock-Out, Standard Deviation, Expressing, Negative Control